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首页> 外文期刊>The American Journal of Tropical Medicine and Hygiene >Comparison of Nested Polymerase Chain Reaction and Real-Time Polymerase Chain Reaction with Parasitological Methods for Detection of Strongyloides stercoralis in Human Fecal Samples
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Comparison of Nested Polymerase Chain Reaction and Real-Time Polymerase Chain Reaction with Parasitological Methods for Detection of Strongyloides stercoralis in Human Fecal Samples

机译：巢式聚合酶链反应和实时聚合酶链反应与寄生虫方法检测人粪便样品中甾体类固醇的比较

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摘要

This study was performed to evaluate nested polymerase chain reaction (PCR) and real-time PCR methods for detection of Strongyloides stercoralis in fecal samples compared with parasitological methods. A total of 466 stool samples were examined by conventional parasitological methods (formalin ether concentration [EEC] and agar plate culture [APC). DNA was extracted using an in-house method, and mitochondrial cytochrome c oxidase subunit 1 and 18S ribosomal genes were amplified by nested PCR and real-time PCR, respectively. Among 466 samples, 12.7% and 18.2% were found infected with S. stercoralis by FEC and APC, respectively. DNA of S. stercoralis was detected in 18.9% and 25.1% of samples by real-time PCR and nested PCR, respectively. Considering parasitological methods as the diagnostic gold standard, the sensitivity and specificity of nested PCR were 100% and 91.6%, respectively, and that of real-time PCR were 84.7% and 95.8%, respectively. However, considering sequence analyzes of the selected nested PCR products, the specificity of nested PCR is increased. In general, molecular methods were superior to parasitological methods. They were more sensitive and more reliable in detection of S. stercoralis in comparison with parasitological methods. Between the two molecular methods, the sensitivity of nested PCR was higher than real-time PCR.
机译：与寄生虫学方法相比，这项研究的目的是评估巢式聚合酶链反应（PCR）和实时PCR方法检测粪便样品中甾体类固醇的能力。通过常规的寄生虫学方法（福尔马林醚浓度[EEC]和琼脂平板培养[APC]）检查了466个粪便样品。使用内部方法提取DNA，并通过巢式PCR和实时PCR分别扩增线粒体细胞色素C氧化酶亚基1和18S核糖体基因。在466个样本中，FEC和APC分别发现了12.7％和18.2％的固醇链球菌感染。实时荧光定量PCR和巢式PCR分别检测到固相链球菌的DNA占18.9％和25.1％。以寄生虫学方法为诊断金标准，巢式PCR的敏感性和特异性分别为100％和91.6％，实时PCR的敏感性和特异性分别为84.7％和95.8％。但是，考虑到所选嵌套式PCR产物的序列分析，嵌套式PCR的特异性增加了。通常，分子方法优于寄生虫学方法。与寄生虫学方法相比，它们在检测S. stercoralis时更灵敏，更可靠。在这两种分子方法之间，巢式PCR的灵敏度高于实时PCR。

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